Image of the Month!

Single Plane Illumination Microscopy

A Single Plane Illumination Microscope (SPIM) is being developed at CABI to
record high-resolution, three-dimensional, optical images. Constructed with two
lasers, this system will allow illumination at two different wavelengths. The
system is being built following the OpenSPIM specification, in collaboration with Pavel Tomancak at the Max Planck Institute of
Molecular Cell Biology and Genetics (MPI-CBG), Dresden.

SPIM principle

In SPIM, optical sectioning is achieved by illuminating the
sample with a sheet of laser light. The specimen is attached to a stage,
rotated and translated orthogonally to the light sheet. Image stacks are
acquired along multiple directions and then merged into a single
three-dimensional data set. The specimen is illuminated from one side and
fluorescent light is collected by a detector positioned perpendicularly to the
light sheet. Since each plane is illuminated only once, fluorophores outside
the focal plane are not exposed to light, and photo damage is therefore
dramatically reduced.

Figure 1. SPIM schematic.

Figure 2. OpenSPIM rendering with laser on, MPI-CBG

SPIM applications

SPIM is a very promising imaging technique that allows the
registration of single-cell resolution images of various kinds of samples.
Indeed, it can not only be used to examine large cleared specimens, but also to
study the development of small insects and animals (drosophila, zebra fish)
over several days.

Video 1. His-YFP drosophila embryo imaged from gastrulation until
muscle contractions in late embryogenesis - ventral view. Used with the permission from Pavel Tomancak and Peter
Gabriel Pitrone, MPI-CBG.